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HPCE法与HPLC法用于甘草中主要活性成分甘草酸及其降解产物甘草次酸分析的比较
Comparison between HPCE and HPLC for the analysis of main active components in licorice
【摘要】 目的:系统地比较高效毛细管电泳法(HPCE)和高效液相色谱法(HPLC)对甘草药材水提取物中主要活性成分甘草酸及其降解产物甘草次酸的分离测定结果。方法:以40 mmol·L-1硼酸盐溶液(pH=9.2)为电泳介质,未涂层弹性石英毛细管(75μm×47.5 cm,有效分离长度40 cm)为分离通道,压力进样(250 kPa·s),20 kV 恒压电泳(25℃)分离,254 nm 检测,内标法定量;HPLC 色谱条件:Beckman C18柱(5 μm,4.6 min×250 mm),流速:0.8 mL·min-1;检测波长:254 nm;流动相 A:水(0.05%三氟乙酸,v/v)、B:乙腈(0.05%三氟乙酸,v/v);梯度洗脱。结果:HPCE 与 HPLC 对甘草酸含量的线性范围分别为0~500μg·mL-1(r=0.9997)、0~1000μg·mL-1(r=0.9999);HPCE 法测得甘草酸平均回收率为102.9%,RSD 为0.5%;HPLC 法测得甘草酸平均回收率为98.8%,RSD 为0.4%。结论:通过对甘草酸分析时间、检出限、线性范围、精密度、重现性、相关性等参数的系统比较,发现 HPCE 法与 HPLC 法对不同种类甘草中甘草酸含量的测定结果有较好的相关性,并且验证了HPCE 同时检测甘草酸及其降解产物甘草次酸的可行性。
【Abstract】 Objective:To compare an HPCE and an HPLC methods for separating and determining the bioactive compound glycyrrhizinic acid (GA) in aqueous extract of licorice,the root of Glycyrrhiza uralensis Fisch.Also to es- tablish an HPCE method for determining GA and its degradation product glycyrrhetinic acid(GTA) simultaneously. Methods:The dimensions of the capillary tube used in HPCE analysis were 47.5 cm×75 μm and the effective length was 40 cm.The temperature of the capillary tube was maintained at 25 ℃.The buffer solution contained 40 mmol·L-1 sodium borate at a pH of 9.2.All CE runs were made at an applied voltage of 20 kV,and with a UV de- tection wavelength of 254 nm.The contents of GA and GTA were measured based on internal standards.The HPLC analyses were performed on a Beckman C18 column(5 μm,4.6 mm×250 mm).The mobile phase contained H2O (0.05%TFA,v/v) and CH3CN(0.05%TFA,v/v),and the flow rate was 0.8 mL·min-1 with 254 nm as the de- tecting wavelength.Results:HPCE and HPLC methods were compared for the qualitative and quantitative analysis of GA in the aqueous extracts of licorice.The calibration curve showed good linearity in the range of 0-500 μg· mL-1 (r=0.9997) by HPCE and of 0-1000 μg·mL-1 (r=0.9999) by HPLC.The average recoveries of HPCE method and HPLC method were 102.9%and 98.8%,RSDs were 0.5%and 0.4%.Conclusion:The primary ad- vantages of the HPCE method,when compared with HPLC,include the much shorter analysis time and the ability to determine simultaneously GA together with its degradation product GTA.The analytical dynamic range and sensitivi- ty of both methods are comparable.Both of these two methods are attractive techniques for the analysis of traditional Chinese medicines.
【Key words】 HPCE; HPLC; licorice; glycyrrhizinic acid; glycyrrhetinic acid;
- 【文献出处】 药物分析杂志 ,Chinese Journal of Pharmaceutical Analysis , 编辑部邮箱 ,2006年04期
- 【分类号】R284
- 【被引频次】22
- 【下载频次】667